Antimicrobial photodynamic therapeutic effects of cationic amino acid-porphyrin conjugate 4i on Porphyromonas gingivalis in vitro.

BACKGROUND: Porphyromonas gingivalis (P. gingivalis) is considered to be among the principal pathogens in periodontal disease. The present study aimed to investigate the effect of antimicrobial photodynamic therapy (aPDT) mediated by cationic amino acid-porphyrin conjugate 4i on P. gingivalis METHODS: The uptake of 4i by P. gingivalis over different times of incubation was evaluated by optical density using a microplate reader. Laser radiation at λ=650nm-660nm with I =50 mW/cm2 at doses of 0, 3.0, 6.0, 9.0, and 12 J/cm2 was used for aPDT. A colony-counting method and confocal laser scanning microscopy (CLSM) were used to observe the neutralization of P. gingivalis. The fluorescent molecular probe 3'(p-hydroxyphenyl)-fluorescein and the reagent Singlet Oxygen Sensor Green were used to measure the quantities of •OH and 1O2 produced by 4i after irradiation with different light energies.

RESULTS: The 4i conjugate was absorbed gradually by P. gingivalis, reaching a maximum at 30 min. A clear cytotoxic effect on P. gingivalis was observed with aPDT using 62.5 µM 4i, with colony counts dropping by a factor of 3.35 log10, indicating a sterilization rate of 99.95%. Light irradiation resulted principally in the production of • OHby 4i. A live/dead viability assay demonstrated substantial red fluorescence in P. gingivalis treated with aPDT.

CONCLUSIONS: The results suggest that 4i-aPDT caused substantial cytotoxicity in P. gingivalis.